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. Author manuscript; available in PMC: 2018 Jun 30.
Published in final edited form as: J Mol Biol. 2016 Oct 11;429(13):2055–2065. doi: 10.1016/j.jmb.2016.10.004

Figure 6.

Figure 6

MLL4 regulates osteogenic differentiation of C2C12. (A) Schematic of CRISPR knockout of histone H3K4 methyltransferase (KMT). KMT knockout C2C12 cells were differentiated into osteoblast and the differentiation was evaluated by ALP. Dox, doxycycline; ALP, alkaline phosphatase. (B) ALP activity was detected and calculated as in Figure 1B. Each bar indicates the average of two replicates. (C) Quantitative RT-PCR analysis of osteogenic genes in MLL4 knockout C2C12. Myoblast were cultured in growth medium. The medium was replaced with osteoblast differentiation medium containing 100 ng/ml BMP-4 when cells were 50% confluent and cells were differentiated into osteoblast for three days. GFP sgRNA was used as control. Error bars indicate SEM from three replicates. Data is a representative of two independent experiment. (D) ChIP-qPCR analysis with indicated antibody at the Runx2 enhancer in MLL4 knockout C2C12 osteoblast. Cells were differentiated in osteoblast differentiation medium. Primers used for qPCR are indicated in Figure 5A. Results are shown as % signal/input. Error bars indicate SEM from four replicates of qPCR. Data is a representative of two independent experiment.